FLIVO® (FLuorescence in vIVO) is a powerful method for non-invasive detection of caspase activity in vivo. NIR-FLIVO 690 Tracers are non-cytotoxic, cell-permeant fluorescent inhibitors of caspases optimized for use in whole live animals. ICT’s NIR-FLIVO 690 Tracer probe contains the preferred binding sequence for most caspases (Val-Ala-Asp or VAD). This preferred poly caspase tripeptide binding sequence is labeled with *Dylight® 690 dye and a fluoromethyl ketone (FMK) reactive entity. FLIVO® is a registered trademark of ICT. *Dylight® is a registered trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
- Prepare samples and controls.
- Dilute 10X Injection Buffer 1:10 with 45 mL diH20.
- Reconstitute NIR-FLIVO 690 Tracer with 50 μL DMSO.
- Dilute NIR-FLIVO 1:10 with 450 μL 1X Injection Buffer.
- Inject 50 μL intravenously.
- Let NIR-FLIVO circulate for 4 hours.
- Image the animal. NIR-FLIVO 690 Tracer excites at 690 nm and emits at 709 nm.
- If not viewing directly, excise tissue and process samples for further analysis.
- Optional: label excised samples with additional stains, such as Hoechst 33342, or an antibody.
- If desired, fix excised samples.
- Analyze excised samples with a fluorescence microscope or flow cytometer.
FLIVO® is a registered trademark of ICT. *Dylight® is a registered trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.